Sintered ferrous amino acid particles and use of the same against a virus

ABSTRACT

A composition includes sintered ferrous amino acid particles prepared by sintering a ferrous amino acid chelate which includes ferrous ions and an amino acid. The sintered ferrous amino acid particles have an average particle size ranging from 500 to 2600 nm and a weight average molecular weight ranging from 1,500 Dalton to 600,000 Dalton. Also disclosed herein are a method for inhibiting and/or killing a virus in a subject and applications of such method. The method includes administering to the subject the composition.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority of U.S. Provisional Patent ApplicationNo. 62/639,064, filed on Mar. 6, 2018.

FIELD

The disclosure relates to a composition including sintered ferrous aminoacid particles, and a method for inhibiting and/or killing a virus usingthe composition, as well as applications thereof.

BACKGROUND

The applicant's US Patent Application Publication No. 2017/0224727 A1has disclosed a ferrous amino acid chelate, which is capable of stablypassing through stomach, and which is effective in controlling bodyweight and enhancing lipid metabolism and lipolysis.

In addition, the ferrous amino acid chelate can also be used in thetreatment of cancer and diabetes, as well as to reduce the production oflactic acid by cancer cells, as disclosed in the applicant's previouspatent applications and patent, including US Patent ApplicationPublication Nos. 2015/0065569 A1 and 2017/0007568 A1 and TaiwaneseInvention Patent No. 1587856. These patent applications and patent arehereby incorporated by reference in their entirety.

Porcine epidemic diarrhea virus (PEDV) is a coronavirus that infects thecells lining the small intestine of a pig, causing porcine epidemicdiarrhoea, a condition of severe diarrhea and dehydration. Older hogsmostly get sick and lose weight after being infected, whereas newbornpiglets usually die within five days of contracting the virus. PEDV hasa substantial economic burden given that it is highly infectious,resulting in significant morbidity and mortality in piglets.

Viral respiratory diseases are also problematic to swine. Among suchviruses, porcine reproductive and respiratory syndrome virus (PRRSV) andswine influenza virus (SIV) are the two main known contributors to lunginfectious diseases, and can take effect alone or in combination.Porcine reproductive and respiratory syndrome virus causes porcinereproductive and respiratory syndrome (PRRS), also known as blue-ear pigdisease, which causes reproductive failure in breeding stock andrespiratory tract illness in young pigs. Swine influenza virus (SIV)causes swine influenza, an acute infectious respiratory disease in pigs,which is characterized by sudden onset, cough, dyspnea, fever and rapidprognosis. Swine influenza has high incidence during autumn and winterthough it can be transmitted all year long. Therefore, there is a needto develop an effective and efficient method of killing the aforesaidinfectious viruses.

The applicant has surprisingly found that the sintered particlesobtained from the ferrous amino acid chelate is effective in inhibitingand/or eliminating a virus, and hence opines that such particles can beused to inhibit and/or eliminate a virus, particularly a porcine virus.

SUMMARY

Accordingly, the present disclosure provides a composition includingsintered ferrous amino acid particles, which are prepared by sintering aferrous amino acid chelate including ferrous ions and an amino acid. Thesintered ferrous amino acid particles have an average particle sizeranging from 500 to 2600 nm and a weight average molecular weightranging from 1,500 Dalton to 600,000 Dalton.

The present disclosure also provides a method for inhibiting and/orkilling a virus in a subject, which includes administering to thesubject the aforementioned composition.

DETAILED DESCRIPTION

Unless defined otherwise, all technical and scientific terms used hereinhave the meaning commonly understood by a person skilled in the art towhich the present disclosure belongs.

One skilled in the art will recognize many methods and materials similaror equivalent to those described herein, which could be used in thepractice of the present disclosure. Indeed, the present disclosure is inno way limited to the methods and materials described. For clarity, thefollowing definitions are used herein.

The present disclosure provides a composition including sintered ferrousamino acid particles, which are prepared by sintering a ferrous aminoacid chelate including ferrous ions and an amino acid. The sinteredferrous amino acid particles have an average particle size ranging from500 to 2600 nm and a weight average molecular weight ranging from 1,500Dalton to 600,000 Dalton.

In certain embodiments, the sintered ferrous amino acid particles have aweight average molecular weight ranging from 1,500 Dalton to 15,000Dalton. In other embodiments, the sintered ferrous amino acid particleshave a weight average molecular weight ranging from 400,000 Dalton to550,000 Dalton. In an exemplary embodiment, the sintered ferrous aminoacid particles have a weight average molecular weight of about 550,000Dalton.

According to this disclosure, the chelating ratio of the ferrous ions tothe amino acid in the ferrous amino acid chelate ranges from 1:1 to 1:4.In certain embodiments, the chelating ratio of the ferrous ions to theamino acid in the ferrous amino acid chelate ranges from 1:1.5 and1:2.5.

The process for preparing the ferrous amino acid chelate has beendisclosed in, e.g. US 2017/0224727 A1, and includes the steps of mixinga ferrous compound with an amino acid under heating. In certainembodiments, the mixing step may be conducted at a temperature rangingfrom 60° C. to 90° C. In certain embodiments, the mixing step may beconducted for 8 hours to 48 hours.

According to the disclosure, the weight ratio of the ferrous compoundand the amino acid used in the preparation process is between 1:1.2 and1:1.5. In an embodiment of this disclosure, the weight ratio of theferrous compound and the amino acid is 1:1.3.

In certain embodiments, the ferrous compound may be ferrous sulfate,ferrous chloride, ferrous pyrophosphate, or the combinations thereof.

In certain embodiments, the amino acid may be glycine. That is, theferrous amino acid chelate may be a ferrous glycinate chelate.

The present disclosure also provides a method for inhibiting and/orkilling a virus in a subject, which includes administering to thesubject the aforementioned composition.

Examples of the virus suitable for use in this disclosure include, butare not limited to, porcine epidemic diarrhea virus, porcine respiratoryand reproductive syndrome virus, influenza virus (such as swineinfluenza virus, canine influenza virus, equine influenza virus, avianinfluenza virus, etc.), transmissible gastroenteritis coronavirus,feline coronavirus, canine coronavirus, equine arteritis virus, andcombinations thereof.

The composition according to this disclosure may be prepared in the formof a pharmaceutical composition or a food composition.

If the composition is prepared in the form of the pharmaceuticalcomposition, the composition may further include a pharmaceuticallyacceptable carrier, and may be made into a dosage form suitable for oraladministration using technology well-known to those skilled in the art.Examples of the dosage form include, but are not limited to, solution,suspension, emulsion, powder, tablet, pill, syrup, lozenge, troche,chewing gum, capsule, slurry and the like.

Examples of the pharmaceutically acceptable carrier suitable for use inthis disclosure may include, but are not limited to, solvent,emulsifier, suspending agents, decomposers, binding agents, excipients,stabilizing agents, chelating agents, diluents, gelling agents,preservatives, lubricants, absorption delaying agents, liposomes, andcombinations thereof.

The composition according to this disclosure may be in the form of afood additive (an exemplary example of the food composition), which canbe added into an edible material to prepare a food product for human oranimal consumption. Examples of the food product according to thisdisclosure may include, but are not limited to: fluid milk products,e.g., milk and concentrated milk; fermented milk, e.g., yogurt, sourmilk and frozen yogurt; milk powder; ice cream; cream cheeses; drycheeses; soybean milk; fermented soybean milk; vegetable-fruit juices;fruit juices; sports drinks; confectionery; jelly; candies; healthfoods; animal feeds; and dietary supplements.

As used herein, the term “subject” refers to any animal of interest,such as humans, monkeys, cows, sheeps, horses, pigs, goats, dogs, cats,mice and rats. In certain embodiments, the subject may be a human. Inother embodiments, the subject may be a pig.

The dosage and the frequency of administration of the compositionaccording to this disclosure may vary depending on the followingfactors: the severity of the virus to be inhibited and/or killed, andthe weight, age, physical condition and response of the subject to betreated. For instance, the daily dosage of the composition according tothis disclosure may be 12 to 36 mg per kg of the body weight, and may beadministered in a single dose or in several doses.

This disclosure will be further described by way of the followingexamples. However, it should be understood that the following examplesare solely intended for the purpose of illustration and should not beconstrued as limiting the disclosure in practice.

EXAMPLES General Experimental Materials: 1. Pharmaceutical CompositionA1:

Pharmaceutical Composition A1 (batch number: F171001; production date:Oct. 5, 2017), which contained sintered ferrous amino acid particles,was in the form of lyophilized powder, and was produced by TaiwanBioligand Co., Ltd.

Specifically, ferrous sulfate was mixed with glycine (above 98% purity)in a weight ratio of 1:1.3, followed by heating from 60° C. to 90° C.for 8 hours to 48 hours to obtain ferrous amino acid chelates with achelating ratio of the ferrous irons to the amino acid ranging from 1:1to 1:4. The ferrous amino acid chelates were then sintered at atemperature ranging from 200° C. to 240° C. to obtain the sinteredferrous amino acid particles.

The average particle size of the sintered ferrous amino acid particlesmeasured in water by dynamic light scattering (DLS) on Beckman CoulterN5 Submicron Particle Size Analyzer was 1465.90±132.29 nm.

The number-average molecular weight (Mn), weight-average molecularweight (Mw), peak molecular weight (MP) and polydispersity (PDI) of thesintered ferrous amino acid particles dissolved in water, determined bygel permeation chromatography using Waters Alliance 2695 System, were68188 Dalton, 525538 Dalton, 286426 Dalton and 7.707205, respectively.

Pharmaceutical Composition A1 was dissolved in sterile purified water toprepare a stock solution having a concentration of 250 mg/mL for use inthe following examples.

2. Virus:

Porcine epidemic diarrhea virus (PEDV), porcine respiratory andreproductive syndrome virus (PRRSV) and swine influenza virus (SIV) wereobtained from Shanghai Academy of Agricultural Sciences (SAAS), China.

3. Host Cells for Propagating Viruses:

Vero cells (ATCC CCL-81) for propagating PEDV, MARC 145 cells (ATCCCRL-12231) for propagating PRRSV, and MDCK cells (ATCC CCL-34) forpropagating SIV were obtained from Shanghai Academy of AgriculturalSciences (SAAS), China.

Example 1. Effect of Pharmaceutical Composition A1 on Porcine EpidemicDiarrhea Virus (PEDV) Experimental Procedures:

A. Pretreatment of Host Cells with Pharmaceutical Composition A1 BeforeVirus Inoculation, and Subsequent Virus Inoculation of Pretreated HostCells with PEDV

Well grown Vero cells, after detachment, were subjected to dilutionusing a DMED medium so as to reach a cell concentration of 10⁶ cell/mL.To each of aliquot portions of the resulting cell suspension containingthe DMED medium was added a suitable amount of the stock solutioncontaining Pharmaceutical Composition A1, so that cell suspensionsrespectively having final concentrations of Pharmaceutical CompositionA1 being 1000 μg/mL, 500 μg/mL, and 100 μg/mL were obtained. On a24-well cell cultivation plate, the cells treated with PharmaceuticalComposition A1 at a respective concentration were seeded into 6 wells,and a cell control (without the treatment of the stock solutioncontaining Pharmaceutical Composition A1) was seeded as well. The cellcultivation plate was placed at 37° C. for 24 hours of incubation, andwas then retrieved to remove the medium via suction. The cells werewashed thrice using a sterile PBS solution.

A PEDV solution was diluted at a ratio of 1:200. 100 μL of the resultingdiluted virus solution was inoculated into each well. Virus adsorptionwas conducted at 37° C. for 1 hour. Afterward, the virus solution wasremoved via suction. 100 μL of a DMEM complete medium was added intoeach well, followed by incubation at 37° C. The cytopathic effect wasobserved. Each of the virus-containing media was collected at 96 hour soas to determine the TCID₅₀ (50% tissue culture infection dose) of thevirus.

B. Virus Inoculation of Host Cells with Pharmaceutical CompositionA1-Treated PEDV

Well grown Vero cells, after detachment, were subjected to dilution soas to reach a cell concentration of 10⁶ cell/mL, followed by cellplating onto a 24-well cell cultivation plate. Incubation was conductedat 37° C. for 24 hours.

A sterile PBS solution was used to dilute PEDV at a ratio of 1:200. Toeach of aliquot portions of the resulting diluted PEDV solution wasadded a suitable amount of the stock solution containing PharmaceuticalComposition A1, so that Pharmaceutical Composition A1-treated PEDVsolutions respectively having final concentrations of PharmaceuticalComposition A1 being 1000 μg/mL, 500 μg/mL, and 100 μg/mL were obtained.The treatment was allowed to proceed at 37° C. for 1 hour.

Subsequently, the Pharmaceutical Composition A1-treated PEDV solutionswere inoculated into the 24-well cell cultivation plate. Specifically, arespective one of the Pharmaceutical Composition A1-treated PEDVsolutions was inoculated into 6 wells, and each well received 100 μL ofthe respective Pharmaceutical Composition A1-treated PEDV solution. Inaddition, a cell control was inoculated virus without the treatment ofPharmaceutical Composition A1.

The cell cultivation plate was placed at 37° C. for incubation. Thecytopathic effect was observed. Each of the virus-containing solutionswas collected at 96 hour so as to determine the TCID₅₀ of the virus.

Results:

A. Pretreatment of Host Cells with Pharmaceutical Composition A1 BeforeVirus Inoculation, and Subsequent Virus Inoculation of Pretreated HostCells with PEDV

The cytopathic effect observed and the TCID₅₀ of the virus determined inthe Vero host cells pretreated with Pharmaceutical Composition A1 areshown in Tables 1 and 2 below, respectively.

TABLE 1 Degree of cytopathic effect observed Concentration ofPharmaceutical Composition A1 1^(st) well 2^(nd) well 3^(rd) well 4^(th)well 1000 μg/mL ++++ ++++ ++++ ++++  500 μg/mL ++++ ++++ ++++ ++++  100μg/mL ++++ ++++ ++++ ++++   0 μg/mL ++++ ++++ ++++ ++++

TABLE 2 TCID₅₀ determined Concentration of Pharmaceutical Composition A1TCID₅₀ 1000 μg/mL 10⁻¹/0.1 mL  500 μg/mL 10^(−1.6667)/0.1 mL  100 μg/mL10^(−2.2241)/0.1 mL   0 μg/mL 10^(−6.3441)/0.1 mL

As shown in Table 2, the higher the concentration of PharmaceuticalComposition A1, the higher the TCID₅₀ determined was, indicating that apretreatment with a suitable concentration of Pharmaceutical CompositionA1 can lead to inhibition against propagation of PEDV in the host cells.

B. Virus Inoculation of Host Cells with Pharmaceutical CompositionA1-Treated PEDV

The cytopathic effect observed and the TCID₅₀ of the PharmaceuticalComposition A1-treated PEDV determined in the host cells are shown inTables 3 and 4 below, respectively.

TABLE 3 Degree of cytopathic effect observed Concentration ofPharmaceutical Composition A1 1^(st) well 2^(nd) well 3^(rd) well 4^(th)well 1000 μg/mL +++ +++ ++ ++  500 μg/mL ++ ++ + ++  100 μg/mL ++ ++++ +   0 μg/mL ++++ ++++ ++++ ++++

TABLE 4 TCID₅₀ determined Concentration of TCID₅₀ PharmaceuticalComposition A1 1000 μg/mL 10^(−3.5714)/0.1 mL  500 μg/mL10^(−4.4099)/0.1 mL  100 μg/mL 10^(−5.4099)/0.1 mL   0 μg/mL10^(−6.3441)/0.1 mL

As shown in Table 3, the treatment with Pharmaceutical Composition A1reduced the degree of cytopathic effect, manifesting that PharmaceuticalComposition A1 is effective in inhibiting and/or killing a virus. Inparticular, when either 500 μg/mL or 100 μg/mL of PharmaceuticalComposition A1 was applied, the cytopathic effect was significantlydelayed, and a significantly lower degree of cytopathic effect wasobserved.

As shown in Table 4, the higher the concentration of PharmaceuticalComposition A1, the higher the TCID₅₀ was, indicating that a treatmentwith a suitable concentration of Pharmaceutical Composition A1 can leadto inhibition against propagation of PEDV in the host cells.

Example 2. Effect of Pharmaceutical Composition A1 on PorcineRespiratory and Reproductive Syndrome Virus (PRRSV) ExperimentalProcedures:

A. Pretreatment of Host Cells with Pharmaceutical Composition A1 BeforeVirus Inoculation, and Subsequent Virus Inoculation of Pretreated HostCells with PRRSV

On a 24-well cell cultivation plate, MARC 145 host cells pretreated withPharmaceutical Composition A1 at a respective concentration of 1000μg/mL, 500 μg/mL, and 100 μg/mL, which were prepared according to theprocedure described in section A of Example 1, were seeded into 6 wells,and a cell control (without the treatment of Pharmaceutical CompositionA1) was seeded as well. The cell cultivation plate was placed at 37° C.for 24 hours of incubation, and was then retrieved to remove the mediumvia suction. The cells were washed thrice using a sterile PBS solution.

A PRRSV solution was diluted at a ratio of 1:200. 100 μL of theresulting diluted virus solution was inoculated into each well. Virusadsorption was conducted at 37° C. for 1 hour. Afterward, the virussolution was removed via suction. 100 μL of a DMEM complete medium wasadded into each well, followed by incubation at 37° C. Each of thevirus-containing media was collected at 92 hour so as to determine thevirus contents in the host cells pretreated with differentconcentrations of Pharmaceutical Composition A1, relative to the cellcontrol.

B. Virus Inoculation of Host Cells with Pharmaceutical CompositionA1-Treated PRRSV

Pharmaceutical Composition A1 was dissolved in sterile purified water toprepare a stock solution having a concentration of 250 mg/mL. Well grownMRAC 145 cells, after detachment, were subjected to dilution so as toreach a cell concentration of 10⁶ cell/mL, followed by cell plating ontoa 24-well cell cultivation plate. Incubation was conducted at 37° C. for24 hours.

Pharmaceutical Composition A1-treated PRRSV solutions respectivelyhaving final concentrations of Pharmaceutical Composition A1 being 1000μg/mL, 500 μg/mL, and 100 μg/mL were prepared according to the proceduredescribed in section B of Example 1.

Subsequently, the Pharmaceutical Composition A1-treated PRRSV solutionswere inoculated into the 24-well cell cultivation plate. Specifically, arespective one of the Pharmaceutical Composition A1-treated PRRSVsolutions was inoculated into 6 wells, and each well received 100 μL ofthe respective Pharmaceutical Composition A1-treated PRRSV solution. Inaddition, a cell control was inoculated virus without the treatment ofPharmaceutical Composition A1.

The cell cultivation plate was placed at 37° C. for incubation. Each ofthe virus-containing solutions was collected at 96 hour so as todetermine the virus contents in the cells inoculated with differentconcentrations of Pharmaceutical Composition A1-treated PRRSV solutions,relative to the normal cell control.

Results:

A. Pretreatment of Host Cells with Pharmaceutical Composition A1 BeforeVirus Inoculation, and Subsequent Virus Inoculation of Pretreated HostCells with PRRSV

The virus content determined in the host cells pretreated withPharmaceutical Composition A1 is shown in Table 5.

TABLE 5 Virus content determined in the host cells Concentration ofVirus content Pharmaceutical (relative to Composition A1 the cellcontrol) 1000 μg/mL 0.322  500 μg/mL 0.664  100 μg/mL 0.876   0 μg/mL 1

As shown in Table 5, the higher the concentration of PharmaceuticalComposition A1, the lower the virus content was, indicating that apretreatment with a suitable concentration of Pharmaceutical CompositionA1 can lead to inhibition against propagation of PRRSV in the hostcells.

B. Virus Inoculation of Host Cells with Pharmaceutical CompositionA1-Treated PRRSV

The virus content determined in the host cells inoculated withPharmaceutical Composition A1-treated PRRSV is shown in Table 6 below.

TABLE 6 Virus content determined in the host cells Concentration ofVirus content Pharmaceutical (relative to the Composition A1 normal cellcontrol) 1000 μg/mL 0.681  500 μg/mL 0.773  100 μg/mL 0.96   0 μg/mL 1

As shown in Table 6, the treatment with Pharmaceutical Composition A1reduced the virus content in the host cells, manifesting thatPharmaceutical Composition A1 is effective in inhibiting and/or killinga virus. In particular, the higher the concentration of PharmaceuticalComposition A1, the lower the virus content was, indicating that atreatment with a suitable concentration of Pharmaceutical Composition A1can lead to inhibition against propagation of PRRSV in the host cells.

Example 3. Effect of Pharmaceutical Composition A1 on Swine InfluenzaVirus (SIV) Experimental Procedures:

A. Pretreatment of Host Cells with Pharmaceutical Composition A1 BeforeVirus Inoculation, and Subsequent Virus Inoculation of Pretreated HostCells with SIV

On a 24-well cell cultivation plate, MDCK cells treated withPharmaceutical Composition A1 at a respective concentration of 1000μg/mL, 500 μg/mL, and 100 μg/mL, which were prepared according to theprocedure described in section A of Example 1, were seeded into 6 wells,and a cell control (without the treatment of Pharmaceutical CompositionA1) was seeded as well. The cell cultivation plate was placed at 37° C.for 24 hours of incubation, and was then retrieved to remove the mediumvia suction. The cells were washed thrice using a sterile PBS solution.

A SIV solution was diluted at a ratio of 1:200. 100 μL of the resultingdiluted virus solution was inoculated into each well. Virus adsorptionwas conducted at 37° C. for 1 hour. Afterward, the virus solution wasremoved via suction. 100 μL of a serum-containing complete medium wasadded into each well, followed by incubation at 37° C. Each of thevirus-containing media was collected at 72 hour so as to determine theTCID₅₀ of the virus using the Reed-Muench method.

B. Virus Inoculation of Host Cells with Pharmaceutical CompositionA1-Treated SIV

Pharmaceutical Composition A1 was dissolved in sterile purified water toprepare a stock solution having a concentration of 250 mg/mL. Well grownMDCK cells, after detachment, were subjected to dilution so as to reacha cell concentration of 10 cell/mL, followed by cell plating onto a24-well cell cultivation plate. Incubation was conducted at 37° C. for24 hours. Pharmaceutical Composition A1-treated SIV solutionsrespectively having final concentrations of Pharmaceutical CompositionA1 being 1000 μg/mL, 500 μg/mL, and 100 μg/mL were prepared according tothe procedure described in section B of Example 1.

Subsequently, the Pharmaceutical Composition A1-treated SIV solutionswere inoculated into the 24-well cell cultivation plate. Specifically, arespective one of the Pharmaceutical Composition A1-treated SIVsolutions was inoculated into 6 wells, and each well received 100 μL ofthe respective Pharmaceutical Composition A1-treated SIV solution. Inaddition, a cell control was inoculated virus without the treatment ofPharmaceutical Composition A1.

The cell cultivation plate was placed at 37° C. for incubation. Each ofthe virus-containing solution was collected at 72 hour so as todetermine the TCID₅₀ of the virus using the Reed-Muench method.

Results:

A. Pretreatment of Host Cells with Pharmaceutical Composition A1 BeforeVirus Inoculation, and Subsequent Virus Inoculation of Pretreated HostCells with SIV

The TCID₅₀ of the virus determined in the host cells pretreated withPharmaceutical Composition A1 is shown in Table 7 below.

TABLE 7 TCID₅₀ determined determined in the host cells Concentration ofPharmaceutical Composition A1 TCID₅₀ 1000 μg/mL 10^(−2.7)/0.1 mL  500μg/mL 10^(−3.18)/0.1 mL  100 μg/mL 10^(−3.39)/0.1 mL   0 μg/mL10^(−4.88)/0.1 mL

As shown in Table 7, the higher the concentration of PharmaceuticalComposition A1, the higher the TCID₅, was, indicating that apretreatment with a suitable concentration of Pharmaceutical CompositionA1 can lead to inhibition against propagation of SIV in the host cells.

B. Virus Inoculation of Host Cells with Pharmaceutical CompositionA1-Treated SIV

The TCID₅₀ of the Pharmaceutical Composition A1-treated SIV determinedin the host cells is shown in Table 8 below.

TABLE 8 TCID₅₀ determined in the host cells Concentration ofPharmaceutical Composition A1 TCID₅₀ 1000 μg/mL 10^(−2.28)/0.1 mL  500μg/mL 10^(−3.45)/0.1 mL  100 μg/mL 10^(−3.59)/0.1 mL   0 μg/mL10^(−4.88)/0.1 mL

As shown in Table 8, the treatment with Pharmaceutical Composition A1reduced the virus propagation, manifesting that PharmaceuticalComposition A1 is effective in inhibiting and/or killing a virus. Inparticular, the higher the concentration of Pharmaceutical CompositionA1, the higher the TCID₅₀ was, indicating that a treatment with asuitable concentration of Pharmaceutical Composition A1 can lead toinhibition against propagation of SIV in the host cells.

Based on the above results, the applicant surprisingly found that thesintered ferrous amino acid particles of this disclosure obtained from aferrous amino acid chelate can inhibit and/or kill a virus, therebysignificantly reducing the virus propagation in host cells.

All patents and literature references cited in the present specificationas well as the references described therein, are hereby incorporated byreference in their entirety. In case of conflict, the presentdescription, including definitions, will prevail.

In the description above, for the purposes of explanation, numerousspecific details have been set forth in order to provide a thoroughunderstanding of the embodiments. It will be apparent, however, to oneskilled in the art, that one or more other embodiments may be practicedwithout some of these specific details. It should also be appreciatedthat reference throughout this specification to “one embodiment,” “anembodiment,” an embodiment with an indication of an ordinal number andso forth means that a particular feature, structure, or characteristicmay be included in the practice of the disclosure. It should be furtherappreciated that in the description, various features are sometimesgrouped together in a single embodiment, figure, or description thereoffor the purpose of streamlining the disclosure and aiding in theunderstanding of various inventive aspects, and that one or morefeatures or specific details from one embodiment may be practicedtogether with one or more features or specific details from anotherembodiment, where appropriate, in the practice of the disclosure.

While the disclosure has been described in connection with what areconsidered the exemplary embodiments, it is understood that thisdisclosure is not limited to the disclosed embodiments but is intendedto cover various arrangements included within the spirit and scope ofthe broadest interpretation so as to encompass all such modificationsand equivalent arrangements.

What is claimed is:
 1. A composition comprising sintered ferrous aminoacid particles prepared by sintering a ferrous amino acid chelate whichincludes ferrous ions and an amino acid, the sintered ferrous amino acidparticles having an average particle size ranging from 500 to 2600 nmand having a weight average molecular weight ranging from 1,500 Daltonto 600,000 Dalton.
 2. The composition of claim 1, wherein the sinteredferrous amino acid particles have a weight average molecular weightranging from 400,000 Dalton to 600,000 Dalton.
 3. The composition ofclaim 1, wherein the ferrous amino acid chelate is a ferrous glycinatechelate.
 4. The composition of claim 1, wherein the chelating ratio ofthe ferrous ions to the amino acid in the ferrous amino acid chelateranges from 1:1 to 1:4.
 5. A method for inhibiting and/or killing avirus in a subject, the method comprising: administering to the subjecta composition of claim
 1. 6. The method of claim 1, wherein thecomposition is orally administered.
 7. The method of claim 1, whereinthe composition is a pharmaceutical composition.
 8. The method of claim1, wherein the virus is selected from the group consisting of porcineepidemic diarrhea virus, porcine respiratory and reproductive syndromevirus, influenza virus (swine, canine, equine influenza, avian influenzaviruses), transmissible gastroenteritis coronavirus, feline coronavirus,canine coronavirus, equine arteritis virus and combinations thereof.